PD-L1对过敏性鼻炎小鼠Treg/Th17细胞免疫失衡调节作用研究  

The effects of PD-L1 on imbalance of Treg/Th17 cell in allergic rhinitis of a murine model

作　　者：王頔 唐桥斐[1] 姜玉秋[1] 张爽[1] 李赛楠[1] 闫智永[1] Wang Di;Tang Qiaofei;Jiang Yuqiu(Otolaryngology Department,Second Affiliated Hospital,Shenyang Medical College,Shenyang 110200)

机构地区：[1]沈阳医学院附属第二医院耳鼻喉科,沈阳110200

出　　处：《陕西医学杂志》2018年第3期287-291,共5页Shaanxi Medical Journal

基　　金：沈阳医学院科研基金资助项目(20161006)

摘　　要：目的:探讨PD-L1对过敏性鼻炎小鼠Treg/Th17细胞免疫失衡的调节作用。方法:将30只BALB/c小鼠随机分三组,模型组和PD-L1重组蛋白干预组给予卵清蛋白(OVA)诱发过敏性鼻炎,PD-L1重组蛋白干预组给予PD-L1重组蛋白进行处理;正常对照组在相同时间点给予生理盐水。采用叠加量化记分法进行搔鼻、喷嚏及鼻溢症状评价;采用HE染色观察鼻黏膜组织的病理变化,采用免疫组化检测鼻黏膜组织PD-1和PD-L1的表达;Western blot检测鼻粘膜组织pAKT、、AKT、mTOR、p-mTOR的表达;RT-PCR检测鼻黏膜组织调节性T细胞(Treg)和辅助性T细胞17(Th17)重要转录因子Foxp3、RORγt表达;ELISA法检测血清IL-10、IL-17A、IL-6的水平。结果:正常对照组、模型组、PD-L1重组蛋白干预组行为学评分分别为0、(6.33±1.15)、(3.67±0.58)分,组间比较具有统一学意义(P<0.05)。与正常对照组相比较,模型组鼻粘膜组织损伤严重,鼻粘膜组织中PD-1、PD-L1和Foxp3的表达下降,RORγt、p-AKT、p-mTOR的表达增加,血清IL-10水平下降,血清中IL-17A、IL-6的水平升高,AKT、mTOR的表达无明显变化。与模型组相比较,PD-L1重组蛋白干预组鼻粘膜组织损伤减轻,鼻粘膜组织中PD-1和PD-L1和Foxp3的表达上升,RORγt、p-AKT、p-mTOR表达下降,血清IL-10水平升高,血清中IL-17A、IL-6的水平下降;组间比较差异显著(P<0.05);AKT、mTOR无明显变化。结论:PD-L1通过抑制AKT-mTOR信号进而促进Treg的分化抑制过敏性鼻炎的发生和发展。Objective:To investigate the effects of PD-L1 on imbalance of Treg/Th17 cell in allergic rhinitis(AR)of a murine model.Methods:Thirty BALB/c mice were randomly divided into three groups,ten animals in each.The mice in the model group and PD-L1 recombinant protein group(PD-L1 group)received ovalbumin(OVA)to reduce allergic rhinitis.Then,The mice were intervented with recombinant PD-L1 protein.Mice in the control group were administrated normal saline.Pathological findings of nasal mucosa was analyzed by hematoxylin and eosin stain,The positive cells of PD-1 and PD-L1 in nasal mucosa was analyzed by immunohistochemistry.The expression of Foxp3 and RORγt in nasal mucosa was measured by RT-PCR.The concentration of IL-10,IL-6 and IL-17A in serum was detected by ELISA.Results:The behavioral scores for the control,model and PD-L1 groups were 0,(6.33±1.15),(3.67±0.58)respectively(all P<0.05).serious pathological damage occured in nasal mucosa in model group,whereas damage eased in PD-L1 group.Significantly decreased concentration of PD-1,PD-L1,Foxp3 in nasal mucosa and IL-10 in serum was observed in model group compared with that of the control group(P<0.05),while significantly increased expression of IL-17A,IL-6 in serum and p-AKT、p-mTOR RORγt in nasal mucosa was found(P<0.05).AKT and mTOR change non-significantly and the opposite trends for PD-L1 group compared with the model group(P<0.05).Conclusion:PD-L1 may inhibited AR by inhibiting AKT-mTOR signal pathway and then promoting the differentiation of Treg cells.

关 键 词：鼻炎 过敏性 季节性/免疫学 重组蛋白质类 ＠调节性T细胞 TH17细胞 小鼠 

分 类 号：R392.3[医药卫生—免疫学]